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Changes in chondroitin sulfate concentration in rabbit blood plasma depending on the method of its administration.

Glycosaminoglycans were determined in the blood plasma of rabbits as a function of time after intravenous, intramuscular and intragastric administration of chondroitin sulfate preparation. After intravenous administration two phases of chondoitin sulfate elimination from the blood with T1/2 131-135 min and 8.5-9.5 min, respectively were observed. The level of the blood plasma glycosaminoglycans did not increase after intragastric introduction of the chondroitin sulfate preparation. Excretion of glycoaminoglycans in the urine was also determined.

Administration, Oral↗

The pharmacokinetics of cyclosporine. II. Blood plasma distribution and binding studies.

The pharmacokinetic consequences of alterations in the blood plasma distribution of cyclosporine are examined in this study. A series of solutions of cyclosporine of known concentration were prepared in rabbit blood. The plasma, separated at 37 degrees C, was then assayed for cyclosporine using an HPLC method. A mathematical model relating the blood and plasma concentrations of cyclosporine at equilibrium was derived and used to fit the data. The model assumes that the free fraction of the drug in plasma (fp) is constant but that the free fraction in blood is not. In the rabbit, fp was estimated to equal 0.405. This may be attributed to cyclosporine binding to lipoproteins, and to the lower concentrations of lipoproteins in rabbit serum compared to that in human serum. The capacity factor and the binding affinity constant of the drug for blood cells were estimated to equal 3590 micrograms/liter and 0.00682 liter/microgram in the rabbit. These results and the findings of a previous study suggest that the free fraction of the drug in blood increased, while the free intrinsic clearance did not change, with increases in the steady state blood concentration of the drug. It was concluded that the changes in binding characteristics of cyclosporine in blood at higher concentration and not the changes in the metabolic activity of the liver, as measured by intrinsic clearance, are responsible for the nonlinear pharmacokinetic behavior exhibited by cyclosporine in the rabbit.

Animals↗

Prostaglandin F2 alpha in blood plasma and milk of non-pregnant and pregnant buffaloes (Bubalus bubalis).

Radioimmunoassay of prostaglandin F2 alpha (PGF2 alpha) was standardised for sufficient precision, accuracy, sensitivity and specificity. The changes of PGF2 alpha were monitored in peripheral blood plasma and milk of 28 Murrah buffaloes. There were considerable individual variations of PGF2 alpha concentration in blood plasma (27.6 to 110.8 pg/ml) and milk (80.3 to 180.4 pg/ml). A non-significant increase of PGF2 alpha concentration (P greater than 0.05) was observed at the end of the cycle of non-pregnant animals in both fluids but no such increase was observed in pregnant animals. The concentration of PGF2 alpha in milk was significantly higher (P less than 0.01) than that of blood plasma and was positively correlated in non-pregnant (r = 0.738) and pregnant animals (r = 0.385).

Animals↗

Use of relaxation-edited one-dimensional and two dimensional nuclear magnetic resonance spectroscopy to improve detection of small metabolites in blood plasma.

The 1H nuclear magnetic resonance (NMR) spectra of biological samples, such as blood plasma and tissues, are information rich but data complex owing to superposition of the resonances from a multitude of different chemical entities in multiple-phase compartments, hampering detection and subsequent resonance assignments. To overcome these problems, several spectral-editing NMR experiments are described here, combining spin-relaxation filters (based on T(1), T(rho), and T(2)) with both one-dimensional and two-dimensional (2D) NMR spectroscopy. These techniques enable the separation of NMR resonances based on their relaxation times and allow simplification of the complex spectra. In this paper, the approach is exemplified using a control human blood plasma, which is a complex mixture of proteins, lipoproteins, and small-molecule metabolites. In the case of T(1rho)- and T(2)-edited 2D NMR experiments, a "flip-back" pulse was introduced after the relaxation editing to make the phase cycling of the "relaxation filter" and the 2D NMR part independent, thus enabling easy implementation of the phase-sensitive 2D NMR experiments. These methods also permit much higher receiver gains to be used to reduce digitization error, in particular, for the small resonances, which are sometimes vitally important for metabonomics studies. Both pulse sequences and experimental results are discussed for T(1)-, T(1rho)-, and T(2)-filtered COSY, T(2)-filtered phase-sensitive DQF-COSY, and T(1), T(1rho)-, and T(2)-filtered TOCSY NMR.

Blood Chemical Analysis↗

[Chemiluminescence of blood plasma in the presence of hydrogen peroxide].

Chemoluminescence arised after incubation of diluted blood plasma with hydrogen peroxide. A typical pattern of the luminescence development, similar to that obtained on addition of H2O2 to other biological objects, was observed: quick increase and gradual decrease to a steady-state chemoluminescence level. The increase in chemoluminescence, which was noted in some experiments, might be due to elevation in content of both initiators of peroxidation and activators of the chemoluminescence in blood plasma. Kinetics of the chemoluminescence did not correlate with concentration of oxygen in media.

Acute Disease↗

[Proton acceptor properties of the blood plasma of a series of vertebrates].

Comparative studies have been made on the kinetics of thermal denaturation of the blood plasma of various vertebrates (lampreys, teleosts, frogs, tortoises, pigeons, mice, rats, rabbits, cats, dogs, man) by measuring ionization equilibrium of protein solution at elevated temperature. It was demonstrated that during the initial stage of heat denaturation at 58 degrees the blood plasma of all the animals studied binds protons practically linearly. Among the animals studied, the highest protonoacceptor capacity exhibited the plasma of warm-blooded animals; it was lower in tortoises frogs and teleost fishes, being the lowest one in lampreys. Comparison of total electric charge of plasma proteins evaluated by potentiometric titration with data on thermal denturation of plasma revealed positive correlation between the charge and protonoacceptor properties of the plasma.

Animals↗

Change in opioid peptide level in the heart and blood plasma during acute myocardial ischaemia complicated by ventricular fibrillation.

1. The influence of acute myocardial ischaemia (AMI) complicated by ventricular fibrillation (VF) on opioid peptide level in myocardium and blood plasma of rats has been studied. 2. Leu-enkephalin level in myocardium of rats with AMI and VF has been found to be significantly lower than in animals with AMI but without VF. 3. Met-enkephalin level has been found to be significantly increased in both animals with VF and without it. We have not found a significant difference in met-enkephalin level in myocardium of animals with VF and without it. 4. AMI was induced to increase the enkephalin and beta-endorphin level in blood plasma of all the animals, whether VF had occurred or not. 5. Preliminary administration of D-Ala2,Leu5,Arg6-enkephalin, a synthetic analogue of leu-enkephalin, has prevented a decrease of ventricular fibrillation threshold in experimental coronary occlusion. 6. The obtained results allow us to conclude that enkephalins of myocardium but not opioid peptides of blood plasma play an important role in VF occurrence.

Animals↗

[Fast and effective method of extracting and separating progesterone from blood plasma].

A qualitatively new method is proposed for extraction and separation of progesterone from blood plasms, substituting the imported reagent hexane for the homeoproduced diethyl ether and the separating technique via freezing. The technical changes proposed improve the percentage of extraction of progesterone in blood plasma from 80 per cent to 95 per cent, making the method more sensitive to low concentrations of blood plasma progesterone.

Ether↗

[Image and quantity analysis of prostaglandin in rats' blood plasma and Na(+)-K(+)-ATPase in their cerebellum during the prevention of motion sickness by cinnarizine].

To study the mechanism of cinnarizine in preventing motion sickness, TXB2, 6-Keto-PGF1 alpha in rats' blood plasma and Na(+)-K(+)-ATPase activity in the endothelial cells of their cerebellar capillary were measured and analysed by a radioactive immunity analyser and a computer image system. The results showed that TXB2 and 6-Keto-PGF1 alpha in rats' blood plasma in the cinnarizine preventing group (CPG) decreased remarkably, compared with those in the motion sickness group(MSG) (p < 0.05). The activity of Na(+)-K(+)-ATPase in the endothelial cells of rats' cerebellar capillary in CPG was higher than that in MSG (p < 0.01). The authors suggest that the lower concentration of TXB2 and 6-Keto-PGF1 alpha in rats' blood plasma in CPG is closely related to cinnarizine which prevents Ca2+ from entering into the platelets and into the endothelial cells of blood vessels. The higher activity of Na(+)-K(+)-ATPase in the cerebellum may be caused by cinnarizene which dilates the blood vessels in the brain, increases the blood flow therein, and hinders Ca2+ from getting into the cerebellum cells. These change are believed to be the important mechanism of how cinnarizine prevents motion sickness.

Animals↗

[Content of cyclic nucleotides in blood plasma of patients with bronchial asthma].

The ratio of cAMP and cGMP in blood plasma of patients with bronchial asthma was altered depending on the severity of the disease. There was noted a decrease in cAMP content and an increase--in cGMP. Distinct alterations in the cAMP content were not observed in infectious and atopic forms of the asthma. Content of cGMP was higher in blood plasma of patients with infectious form of the asthma as compared with the atopic one. Dynamics of improvement in the patients with the severe type of bronchial asthma was accompanied by the increased in the ratio cAMP/cGMP due to an increase in the cAMP content, while the content of cGMP remained elevated.

Asthma↗

[The use of blood plasma with elevated titers of antibodies to Re-chemotype glycolipid in peritonitis].

The article analyzes the possibility of immunotherapy of septic complications in cases of peritonitis caused by gram-negative bacteria. A strictly inverse correlation between the severity of intoxication and the level of antibodies to glycolipid Re in blood serum has been established. About 5% of healthy nonimmunized donors have elevated Re-antibody titers (1:128 and higher) in their blood plasma. Screening of blood preparations from more than 1000 donors permitted the creation of the blood plasma bank used for the treatment of peritonitis patients. Immunotherapy of such patients has made it possible to decrease almost two-fold the death rate and to reduce the severity and duration of the intoxication syndrome in patients, as well as to improve the results of the treatment of peritonitis.

Acute Disease↗

1H NMR studies on protein binding of histidine, tyrosine and phenylalanine in blood plasma.

The binding of histidine (His), phenylalanine (Phe) and tyrosine (Tyr) to macromolecules in blood plasma and serum has been investigated by high resolution 1H NMR spectroscopy. In single pulse and spin-echo spectra of quality control bovine serum and normal human plasma, there are no resonances in the chemical shift range to high frequency of delta = 5.3 ppm when measured in the pH* (meter reading, uncorrected for the presence of 2H2O) range 3 to 8.5. On acidification of the plasma to pH* less than 2.5, resonances from His and Phe are observable. In plasma from patients with Wilson's disease, weak signals from His and Tyr are seen in spin-echo spectra at pH* 7.6, but increase in strength on acidification and signals from Phe appear at pH* 1.8. Addition of standard solutions containing Tyr, His and especially Phe and Trp to plasma at neutral pH* results in poor recovery of their expected signal intensity in spin-echo spectra. Addition of 2 M urea to bovine plasma at pH* 4.5 results in the appearance of Phe signals. These data are consistent with Phe and Tyr (and to a lesser extent His) being bound in or to a macromolecular structure at neutral pH from which there is relatively slow exchange with the free solution environment. Experiments with model solutions suggest that serum albumin has a high capacity for binding aromatic amino acids (stabilized by hydrophobic interactions) at neutral pH and this is responsible for the NMR-invisibility of Tyr and Phe in blood plasma.

Animals↗

[The virological safety and bacterial sterility of a method for fractionating blood plasma proteins with rivanol].

The bacterial and virological safety of the method of rivanol fractionation of blood plasma proteins has been evaluated in experiments with samples of donor blood plasma mixed with the suspension of viruses and Escherichia coli used as models. The bacteriostatic action of rivanol and the elimination of bacteriophage and influenza virus from the end product at the stages of rivanol precipitation and adsorption on carbon have been established.

Bacteria↗

The determination of magnesium in human blood plasma by 31P magnetic resonance spectroscopy using a macrocyclic reporter ligand.

The ligand 1,4,7-triazacyclononane-1,4,7-tris(methylene methylphosphinic acid), NOTMP, was used to measure free MgII levels in blood plasma by 31P MRS. Separate resonances were observed for the free ligand and the MgII complex and the ratio of their resonance areas was used to evaluate the free, ionized MgII concentration, [Mg]free. The CaII and the ZnII complexes gave rise to separate resonances in the 31P spectrum in an aqueous sample. In human blood plasma samples, however, these resonances were never observed thus excluding the interference of these metal ions. Heparin, up to 150 units/ml, had no influence on the Mg-NOTMP equilibrium. The 31P MRS methodology was applied to twenty human blood plasma samples. Total MgII ([Mg]total), as measured by atomic absorption spectroscopy, averaged 0.85 +/- 0.12 mM while free ionized MgII ([Mg]free) measured by 31P MRS was 0.66 +/- 0.09 mM. The 31P MRS method gave inherently larger values for free ionized MgII than that reported by ion-selective electrodes (ISE). This was traced to a redistribution of existing plasma MgII species after the addition of about 2 mM of NOTMP. Calculations using existing thermodynamic data show that the ionized MgII concentration (iMg) and the concentration of MgII weakly complexed to small anions (Mg(comp)) both drop after the addition of NOTMP, with Mg(comp) dropping to negligible levels. Thus, the 31P MRS method appears to be less sensitive to variations in the concentration of weakly binding anions (bicarbonate, carbonate, chloride, lactate, phosphate, etc.) than the ISE method. Our data indicates that the difference between Mg(total), as measured by atomic absorption spectroscopy, and Mg(free), as measured by 31P MRS, provides an direct estimate of the protein bound MgII fraction.

Anions↗

Optical biosensors for real-time measurement of analytes in blood plasma.

The preparation of assemblies consisting of multiple molecular layers of bovine serum albumin (BSA), monoclonal antibodies against horseradish peroxidase (anti-HRP), and monoclonal antibodies against methotrexate (anti-MTT), as well as interaction of the assemblies with human blood plasma were observed using a grating coupler and Young interferometer (YI). The assemblies could be arranged according to decreasing amounts of nonspecific deposits bound irreversibly to them from blood plasma as follows-an adsorbed antibody monolayer saturated with adsorbed BSA, antibody multilayers linked with polycations, antibodies covalently immobilized on a BSA layer densely crosslinked with glutaraldehyde (GA), slightly crosslinked BSA double layer, slightly crosslinked antibody double layers. The occurrence of human serum albumin (HSA), human fibrinogen (Fg), IgG, and IgM in the plasma deposits was studied by binding the respective antibodies. IgG, IgM, and Fg were detected in plasma deposits on the immobilized assemblies while the composition of a plasma deposit on the unmodified sensor surface reflected roughly the plasma composition containing mainly adsorbed HSA and Fg. A crosslinked anti-HRP double layer was immobilized on a waveguiding branch of YI and a similar anti-MTT double layer was immobilized on the other branch. The sensor response to blood plasma was fairly decreased owing to a compensation of the respective optical changes in the two branches, in which a similar non-specific adsorption took place. The addition of HRP or MTT to plasma induced specific responses of the corresponding branches.

Animals↗

Metal and metalloid multi-elementary ICP-MS validation in whole blood, plasma, urine and hair. Reference values.

Four multi-elementary metal and metalloid quantification methods using inductively coupled plasma mass spectrometry (ICP-MS) were developed and validated in human whole blood, plasma, urine and hair by means of a single preparation procedure for each sample. The ICP-MS measurements were performed using a Thermo Elemental X7CCT series and PlasmaLab software without a dynamic reaction cell. With this procedure 27-32 elements can be simultaneously quantified in biological matrices: Li, Be, B, Al, V, Cr, Mn, Co, Ni, Cu, Zn, Ga, Ge, As, Se, Rb, Sr, Mo, Pd, Ag, Cd, Sn, Sb, Te, Ba, W, Pt, Hg, Tl, Pb, Bi, U. Whole blood, plasma and urine samples (0.4 ml each) were diluted with purified water, acid, triton X100 and butanol. Rhodium was used as internal standard. The urine sample results were corrected for enzymatic creatinine determination. Twenty-five milligrams hair samples were acid mineralized after a decontamination procedure and diluted as previously described for biological fluids. To be validated, each element had to show linearity with a correlation coefficient higher than 0.99. The intra-assay and inter-assay inaccuracy, measured as the variation coefficient, were below 5 and 10% respectively. Global performance was assessed by a quality control program. Our laboratory is a registered participant of the Institut National de Santé Publique du Québec (Sainte-Foy, Canada) inter-laboratory comparison program for whole blood, urine, and beard hair of non-occupationally exposed individuals spiked with selected elements. In our study multi-element metal and metalloid analysis was assessed for 27 elements in whole blood, 27 elements in plasma, 30 elements in urine and 32 elements in hair, from 0 to 25, or 250 to 1000 ng/ml, depending on the element. Quantification limits ranged from 0.002 ng/ml (U) to 8.1 ng/ml (Al) for whole blood, from 0.002 ng/ml (U) to 7.7 ng/ml (Al) for plasma, from 0.001 ng/ml (U) to 2.2 ng/ml (Se) for urine, and from 0.2 pg/mg (Tl) to 0.5 ng/mg (B) for hair. Normal values were determined in whole blood (n=100), plasma (n=100), urine (n=100), and hair (n=45) of healthy volunteers, leading to approximately 10,000 analyses. All results are presented and discussed. Clinical toxicology and forensic toxicology applications are also reported. ICP-MS has made significant advances in the field of clinical biology, particularly in toxicological analysis. This is due to the use of extremely effective equipment that permits better clinical and forensic toxicological analysis of metal and metalloid status of each individual patient.

Aged↗

The determination of nitrite and nitrate in human blood plasma by capillary zone electrophoresis.

The determination of nitrite and nitrate in human blood plasma by capillary zone electrophoresis (CZE) is described. Electrophoresis was carried out in a 65 cm long, 75 microns wide silica capillary, using 750 mM sodium chloride containing 5% NICE-Pak OFM Anion-BT (an additive reversing the electroosmotic flow) as running buffer, at a potential of 20 kV, with on-column UV detection at 214 nm. Under these conditions, the two anions migrated in the order of nitrite and nitrate, completely resolved from each other. Forty-one samples of ultrafiltered blood plasma of healthy volunteers were analyzed. The concentrations of nitrite and nitrate were 0.15 +/- 0.07 mg/L and 3.2 +/- 1.6 mg/L (mean +/- SD), respectively, with a linear correlation in the range of 0.1-50 mg/L. The within-day and between-day coefficients of variation (CVs) for nitrite and nitrate were lower than 5% and 8%, respectively. The recovery was 97-114%. The estimated plasma volume needed for each determination was 108 nL.

Electrophoresis, Capillary↗

[Insulin activity of blood plasma in fetuses and newborn infants with asphyxia. Effect of insulin on the course of hypoxia and elimination of its sequelae under experimental conditions].

In this clinico-experimental work data are presented concerning the blood plasma insulin activity in mature fetuses and neonates in asphyxia; the influence of insulin on the course of hypoxia and elimination of its sequelae under experimental conditions on the newborn and sexually mature rats was also noted. The blood plasma insulin activity proved to increase in the neonates born of mothers with normally coursing pregnancy. A statistically significant reduction of the insulin activity was noted in neonates whose mothers suffered from toxemia of the second half of pregnancy. The death of fetuses and neonates was accompanied by a marked reduction of the blood plasma insulin activity. Preliminary administration of insulin increased the tolerance of experimental animals to hypoxia. Administration of insulin at the period of normalization of physiological functions after hypoxia produced a positive influence on the animal revival. Data are presented on the mechanism of insulin action in hypoxia.

Animals↗